Download What`s in your DNA?

What’s in your DNA?
Name: Sunitha Vege, MS
Manager, Genomics Laboratory, NYBC
Date / Time: Monday, October 8, 2012 / 7:45 AM
Event: AABB Boston Industry Workshop
26 – GLOBCC – 5OCT2014
Background
•
•
•
1850’s:
• G. Mendel – inheritance of
traits;
• “father of genetics”
1950’s:
• Structure of DNA was
discovered
• J. Watson and F. Crick
• R. Franklin and M. Wilkins
1980’s:
• Advent of PCR technology
• K. Mullis
2 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Age of DNA-based Testing
 Biology
 Forensics
 Paternity
 Disease diagnosis
 Microbiology
 Pathology
3 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Evolution of genomics: DNA Extraction
Manual
1-2 hours
12 sample
Semi-automated*
1-2 hours
12 samples
*QIAcube and MDx instruments, QIAGEN, Valencia, CA
4 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Automated*
3 hours
96 samples
Evolution of genomics: PCR testing
Manual assays
Real-time PCR*
Array technologies
DNA probes on:
colored beads
Luminex
miniture beads
on silicone chip
*Eppendorf
Immucor/BioArray
glass slides
hydrostatic
holes
Days/Week
Hours/Days
5 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
<8 hours
Advantage of array technologies
 Type for multiple minor antigens in a single assay
BLOODchip Luminex Assays
*
*
*
*In the US and Canada, the BLOODChip products are for research use only. Not for use in diagnostic procedures.
6 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Current DNA-based testing for patients
 Multiply transfused: DNA isolated from white cells
 +DAT
 Allo versus auto antibodies
 OB patients to determine if candidate for Rh immune
globulin
• RHD screen (weak/partial) to confirm RhD status
- Currently done when serology typing is weaker than expected
- Future: Test all OB patients Determine if candidate for Rh immune
globulin
• Zygosity testing of paternal sample when mother has antibody,
typically to RhD, -K, -c, or HPA-1a
- Type fetus from amniocytes to determine if possibility of hemolytic
disease newborn (HDN) or neonatal alloimmune thrombocytopenia
(NAIT).
7 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Testing of patients with Sickle Cell Disease (SCD)
 Full RH genotype
- RH common - common Rh antigens are:
• D (“Rh positive or Rh negative”)
• C and c, E and e antigens
• Exceptions in African Black ethnic group
-
Many additional Rh antigens (n=>50 antigens)
Not detected with serologic reagents
Have significance for transfusion
Matching program for C, E, and K
• Not truly Rh antigen matched unless genotyped
8 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Current DNA-based testing for donors
 Resolve typing discrepancies such as with Fyb,
ABO, and Rh
 Genotype for antigens for which serologic reagents
are not available or are not reliable
 Screen donors for rare phenotypes
 RH genotyping of donors, particularly of African
Black ethnicity, for potential match for sickle cell
patients with unexpected Rh antibodies
9 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Molecular characterization of panel cells
 Currently, some commercial panel cells (Immucor)
have extended antigen results listed by DNA
testing, i.e. Dombrock
 In specialized reference laboratories, panel cells
characterized for rare RH genotypes for resolving
difficult Rh antibodies
• DIIIa ceS / DIIIa ceS
• r’S / r’S; i.e. (C)ceS
• DAR ceAR / DAR ceAR
10 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Frequently asked questions
 Patient/donor consent:
• not “genetic” testing
• Determination of blood group antigens by different method
• Is a “predicted phenotype”
 Reimbursement for testing:
• CPT codes for molecular testing apply
 Standards:
• AABB
11 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Frequently asked questions
 Not yet FDA approved – research use only – similar to unlicensed
typing sera used in reference lab for typing uncommon antigens
 NYBC disclaimer
• These in vitro diagnostic tests were developed and their performance characteristics established
in the Molecular Analysis Laboratory. The tests have not been submitted to the Food and Drug
Administration (FDA) for clearance or approval and; therefore, are not FDA-licensed tests. The
Molecular Analysis Laboratory is certified under the Clinical Laboratory Improvement Amendment
(CLIA) of 1988 as qualified to perform high complexity clinical testing. The New York Blood Center
has been approved by the New York State Department of Health to perform these tests under its
current Clinical Laboratory Permit.
• These results are intended to predict a blood group antigen profile in a patient or donor, and are
not intended for clinical diagnosis or as the sole means for patient management decisions. There
are situations where testing DNA of a person may not reflect the red cell phenotype. Nucleotide
changes that inactivate gene expression or rare new variant alleles may not be identified in these
assays. Test results obtained from DNA isolated from leukocytes and other hematopoietic cells
may differ from DNA isolated from other tissues in persons with a history of transplantation.
12 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Cost of reagents for serological testing:
Average from several centers
Anti-C
1.15
Anti-E
1.15
Anti-c
1.15
Anti-e
2.20
Anti-K
1.55
Anti-k
11.17
Anti-Kpa
12.13
Anti-Kpb
12.13
Anti-Fya
7.11
Anti-Fyb
11.43
Anti-Jka
12.31
Anti-Jkb
13.21
Anti-S
18.79
Anti-s
8.63
Anti-M
3.56
Anti-N
3.74
Anti-Dia
14.35
Total
$135.76
 Supplies, positive and
negative controls, and
labor significantly
increases this cost
 Total cost: >$150-$180
13 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Goals of DNA-based testing
 Minimize (eliminate) alloimmunization
• Provide more precise matched blood versus “least
incompatible” when patients have warm autoantibodies
 Reduce cost:
• Decrease patient work-ups:
- EGA treatment; hypotonic wash
- Adsorption/elution
• Locating rare blood
• These costs have not been previously calculated but offer
significant savings.
14 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
In the year…
 More personalized health care
 Wikipedia definition:
• Personalized medicine is a medical model that proposes the
customization of healthcare, with decisions and practices
being tailored to the individual patient by use of genetic or
other information*
 Genotype all patients
• Only needs to be done once for a patient
• Results are part of the patient’s electronic medical
record
*Personalized Medicine. (2009, March 16). In Wikipedia, The Free Encyclopedia. Retrieved September 28, 2012,
from http://en.wikipedia.org/w/index.php?title=Personalized_Medicine&oldid=277592839
15 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
Next generation sequencing
 Sequencing of the whole genome
 23 and me
• Submit saliva and get results on
numerous SNPs for only $299
• “Connect to ancestry”
• “Take a more active role in
managing your health”
 Third generation sequencing
• Predicted to sequence whole
genome for a few hundred dollars in
matter of hours.
 Analysis of all the sequence
results is still ongoing
https://www.23andme.com/
16 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only
QUESTIONS
17 | What’s Next? | Sunitha Vege | 8 Oct 2012 | Next-Generation Red Cell Typing | Business Use Only